This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The goal of my project is to understand the metabolic activity that supports microbial growth under extreme starvation. In understanding the interactions and processes that sustain microbial communities in starved environments, we hope to better understand one of the outstanding questions of microbial ecology: why are the majority of organisms identified in the environment unculturable in the laboratory? This unculturability is presumed to be due to the inability to reproduce the fastidious growth requirements of many of these species, making them viable within the environment, but unculturable under laboratory conditions. Through this proposal, we aim to test our hypothesis that microbial communities subsist in extremely oligotrophic environments by establishing mutualistic interactions, allowing species to more efficiently utilize the complex nutrient sources entering the system. While such interactions may promote diversity under extreme starvation, obligate mutualism prevents cultivation of these species using traditional techniques. The experiments proposed are geared toward understanding the role that the amount and type of available energy entering these systems have on community diversity and mutualism. The specific aims are: 1) to determine whether microbial mutualistic interactions are occurring under extreme starvation conditions and what role they play in supporting community growth;2) to determine whether such mutualism increases the usable energy sources and metabolic efficiency under extreme starvation, driving community diversity;and 3) to increase the number of previously unculturable bacterial species from these extremely oligotrophic environments by separating previously interdependent species. By using a combination of techniques to monitor how nutrient flow affects community diversity, we endeavor to understand some of the more complex metabolic interactions capable of supporting life under starvation, while increasing the diversity and number of culturable microbial species.